Metrics details While respiratory adaptation to exercise is compulsory to cope with the increased metabolic demand the neural signals at stake remain poorly identified Using neural circuit tracing and activity interference strategies in mice we uncover here two systems by which the central locomotor network can enable respiratory augmentation in relation to running activity One originates in the mesencephalic locomotor region (MLR) Through direct projections onto the neurons of the preBötzinger complex that generate the inspiratory rhythm the MLR can trigger a moderate increase of respiratory frequency The other is the lumbar enlargement of the spinal cord containing the hindlimb motor circuits and through projections onto the retrotrapezoid nucleus (RTN) it also potently upregulates breathing rate On top of identifying critical underpinnings for respiratory hyperpnea these data also expand the functional implication of cell types and pathways that are typically regarded as “locomotor” or “respiratory” related the underlying cells and circuits are not fully elucidated While this is suggestive of ascending projections to respiratory centers the underlying circuit and its functionality during running has not been documented Here we sought to investigate the central circuits interfacing locomotor and respiratory centers in the resourceful mouse model We found the existence of both a descending drive from the MLR and of an ascending drive from the locomotor CPG of the lumbar spinal cord the MLR is capable of upregulating breathing rate even before the initiation of actual limb movements We further uncovered that the two systems both have access to respiratory rhythm generation mechanisms albeit through two different synaptic schemes while the lumbar spinal cord targets the pF region which in turns contacts the preBötC Our work therefore demonstrates two locomotor central drives that may underlie breathing adaptability during running and their synaptic nodes in the respiratory central network Abbreviations used in all figures: PAG periaqueductal gray Source data are provided as a Source data file these analyses demonstrate that (i) Glut+ CnF neurons can upregulate breathing rate before the highest increase in breathing rate from rest occurs when actual locomotor movements are engaged (iii) respiratory frequency increase during the “loco” phase is proportional to the displacement speed and (iv) breaths are not phase-locked to cyclic limb movements these experiments indicate that the activation of lumbar spinal circuits that contain the locomotor CPG exerts an excitatory effect on respiratory activity These experiments highlight the capacity of spinal lumbar circuits to upregulate respiratory-like activities through RTNPhox2b/Atoh1 neurons in the pF respiratory region which may support a longer latency to locomotor onset c CnF activation eventually leads to a running episode during which ventilation is augmented further This effect is attributed to the activated lCPG sending direct projections to the pF respiratory area and in particular to the RTNPhox2b/Atoh1 neurons which in turns contact and activate the preBötC (thick lines) host projection-defined subsets that each control one trait of a multi-faceted behavior This leaves open the possibility that the two locomotor drives may synergize to set the respiratory frequency during ongoing running Our work strikingly substantiates this view The CnF would pre-empt and trigger heightened metabolic demand at the onset of exercise while the RTN by adjusting the respiratory effort to the engaged and sustained locomotion Although our data are compatible with such a working model the proposed connectivity will need to be investigated directly by future work We provide here a firm demonstration and a circuit characterization of two central neuronal drives for breathing adaptation during running The circuit revealed highlights the multi-functional ambition of cell-types and pathways that are typically regarded as “locomotor” or “respiratory” related This represents an entry point to further decipher the nodes and links through which distinct motor programs necessarily cooperate All animals were kept on the C57BL/6J background and group-housed with free access to food and water in controlled temperature (21 °C) and humidity (between 40 and 55%) conditions and exposed to a conventional 12-h light/dark cycle Experiments were performed on animals of either sex aged 2 to 3 months at the time of first injection and data from males and females were pooled The distribution of males and females for all experimental condition is presented in the Source data files All procedures were approved by the French Ethical Committee (“Comité d’éthique en Expérimentation Animale” authorization 2020-022410231878) and conducted in accordance with EU Directive 2010/63/EU All efforts were made to reduce animal suffering and minimize the number of animals Rachael Neve (Gene Delivery Technology Core the pipette was held in place for 5 to 10 min before being slowly retracted a 200 µm core 0.39 NA optic fiber connected to a 1.25 mm diameter ferrule (Thorlabs) was implanted 0.4 mm above targeted sites Optic fibers were secured to the skull with dental cement (Tetric Evoflow) we injected 600-750 nL of CTB-AF647 conjugate (ThermoFisher Scientific # C-34778) diluted at 0.5% in sterile water on each side of the spinal cord and animals were followed daily after the surgery All animals recovered without motor impairments The protocol is inspired by previous work4 A 12 cm pair of electrodes was prepared from Teflon-coated insulated steel wires with an outside diameter of 0.14 mm (A-M systems the Teflon insulation was stripped over 1 mm from each wire so that the two bare regions were separated by about 2 mm The ends of the two wires were soldered to a miniature dissecting pin and a 5 cm ground wire were soldered to a micro connector (Antelec) Nail polish was used to insulate the wires at the connector Their temperature was maintained at 36 °C with a feedback-controlled heating pad This step was crucial to ensure post-surgery survival The skull was exposed and processed to secure the micro connector using dental cement (Tetric Evofow) The ground wire was inserted under the neck’s skin and the twisted electrodes were tunneled towards the right part of the animal guided by a 10 cm silicon tube of 2 mm inner diameter The animal was then placed in supine position the peritoneum was opened horizontally under the sternum and the silicon tube containing the electrodes was pulled through the opening The sternum was clamped and lifted upwards to expose the diaphragm A piece of stretched sterile parafilm was placed on the upper part of the liver to avoid friction during movement of the animal and to prevent conjunctive tissue formation at the recording sites The miniature dissecting pin was pushed through the right floating ribs The pin was then inserted through the sternum leaving the bare parts of the wires in superficial contact with the diaphragm The electrodes’ position was secured on both sides of floating ribs and sternum using dental cement The pin was removed by cutting above the secured wires The peritoneum and abdominal openings were sutured and a head bar was placed on the cemented skull to facilitate animal’s handling when connecting and disconnecting EMG cables during behavioral sessions Buprenorphine (0.025 mg/kg) was administered subcutaneously for analgesia right after surgery and animals were observed daily following the surgery and treated with Buprenorphine (0.025 mg/kg per day) if needed Adult mice were anesthetized with Euthasol Vet (140 mg/kg) and perfused with 4% paraformaldehyde (PFA) in 1× Phosphate Buffered Saline (PBS) Brains and spinal cords were dissected out and fixed overnight in 4% PFA at 4 °C Brains and spinal cords were cryoprotected overnight at 4 °C respectively in 16% and 20% sucrose in PBS Tissues were rapidly cryo-embedded in OCT mounting medium and sectioned at 30 µm using a cryostat Sections were blocked in a solution of 1× Tris Buffered Saline (TBS) 5% normal donkey serum and 1% Triton X-100 Primary antibodies were detected after 2 h of incubation at room temperature with appropriate affinity-purified secondary antibodies obtained from Jackson ImmunoResearch and used at a final dilution of 1:500: donkey anti-chicken AlexaFluor 488 (ref # 705-605-147) and donkey anti-sheep Cy 3 (ref Sections were counterstained with a fluorescent Nissl stain (NeuroTrace 435/445 blue Thermo Fisher Scientific) and mounted in Prolong Diamond Antifade Montant (P36970 Sections were acquired with a Leica TCS SP8 confocal microscope running the LAS X v3.5 software (NeuroPICT imaging platform of the NeuroPSI Institute) and using ×10 and ×25 objectives or on Zeiss AxioImager running the ZEN 3.4 software using a ×10 objective and exclude an unintentional silencing by over-activation 50 ms light stimulations (50-70 pulses/experiment) were applied randomly in the respiratory cycles customized to allow the passage of the optical patch-cord The plethysmography signal was recorded over a period of 10 min using a National Instruments Acquisition card (USB-6211) and the LabScribe NI v3.0 software (iWorxs) Four to five weeks following the injection of the ChR2-expressing virus in the CnF, animals were implanted with a diaphragm EMG as explained previously (Fig. 3) animals were placed in a linear corridor (80 × 10 cm) and familiarized for 1 h/day for 3 days prior to experiments Implanted animals were filmed from the side at 200 fps and 0.5 ms exposure time using a CMOS camera (Jai GO-2400-USB) and images were streamed to a hard disk using the 2nd LOOK v2.0 software (IO Industries) The start of the EMG recordings was hardware-triggered by the start of the video-recordings using the frame exposure readout of the video camera so that the two recordings are synchronized When animals were immobile at one end of the corridor and their respiration was stable we delivered CnF optogenetic activations with frequencies ranging from 5 to 40 Hz the stimulation was repeated three times with several minutes of rest between trials Animals were placed again on the treadmill with the same paradigm 2-3 h and 5 h after CNO or saline administration to measure respiration in resting and running conditions animals were filmed from the side in the same way as above to monitor the stability of running episodes All values were averaged across animals (3 sections/animal) and a grand mean ± SD across n animals was calculated per hemi-section All values were averaged across animals and a grand mean ± SD across n animals was calculated per hemi-section To assess the rostro-caudal distribution of transfected cells in the spinal cord (Fig. 4b) we used one coronal section from seven segments of the spinal cord: the 12th (T12) and 13th (T13) thoracic and the 1st to 5th lumbar (L1 to L5) segments Cells were counted using ImageJ Cell Counter All values were averaged across animals (1 section/animal) and a grand mean ± SD across n animals was calculated per hemi-section The durations of the respiratory cycle containing the light stimulus (perturbed cycle, θ) and the previous respiratory cycle (control cycle, ɸ, Figs. 2 and 7) were measured One respiratory cycle was defined from the onset of inspiration to the subsequent inspiratory onset The phase of light-stimulation ɸS was defined from the onset of the perturbed cycle to the onset of the light pulse The perturbed cycle θ was defined as from the onset of the inspiration that precedes the light stimulation to the onset of the subsequent inspiration The perturbed phase (phase-shift) was calculated as the ratio of the perturbed cycle divided by the control cycle (θ/ɸ) The light phase was defined as the ratio of the stimulated cycle divided by the control cycle (ɸS/ɸ) The perturbed phase was then plotted against the light phase for all events from all animals A phase shift <1 (perturbed cycle duration lower than the control one) indicates a shortening of the respiratory cycle a phase shift >1 (perturbed cycle duration higher than the control) indicates a lengthening and a phase shift equal to 1 (perturbed cycle duration equal to the control) indicates no effect The number of events (N) and animals (n) are given in the corresponding figures for all tested condition the average perturbed phase was plotted against the average light phase in 0.1 ms bins as mean ± SD averaged for each animal and a grand average was calculated and annotated in the corresponding figures for all tested condition Expiratory time (E) was calculated from respiratory cycle and inspiratory (I) times the head x coordinate was used to calculate the animal’s speed sx using the gradient over time Locomotor onset delay (Fig. S6c) was defined as the latency between the onset of the CnF stimulation and the onset of movement for each CnF stimulations All values were averaged across trials (3 trials/animal) and a grand mean ± SD across n animals was calculated per stimulation frequency For gait analysis during CnF photostimulations (Fig. S6d) we manually annotated the paw of a reference hindlimb (ipsilateral) and registered the timings of footfalls (when the paw first touches down) Each reference locomotor cycle was then defined as the duration from one footfall (ipsi_FFn) to the next (ipsi_FFn+1) The time of occurrence of the contralateral hindlimb footfall within the reference locomotor cycle was annotated manually (contra_FF) and the synchronicity rate was then computed as follows: Ipsilateral and contralateral hindlimb steps were categorized as synchronized (synchronicity rate ∈[0 1]) or alternated (synchronicity rate ∈[0.25 Synchronicity rates were averaged across animals (3 trials/animal) and a grand mean ± SD across n animals was calculated per stimulation frequency Circular plots were obtained using a custom macro in Excel All values were averaged across animals (3 trials/animal) and a grand mean ± SD across n animals was calculated per stimulation frequency during (CNO/saline) and after (REC) administration of either CNO or saline we measured instantaneous respiratory frequency and amplitude using the threshold search in Clampfit Inspiratory (Ti) and expiratory (Te) times were quantified manually before (CTL) These measurements were done using 2 to 3 windows of 6 s each taken during resting conditions and at any stable moment of the 1.5 min run (excluding the first 20 s to avoid possible stress-induced changes when the treadmill is just engaged) Measurements were averaged to give the mean value for each animal Averaged mean values were expressed as mean ± SD across n animals Instantaneous respiratory-like frequencies were analyzed offline using the threshold search in Clampfit (Molecular Devices) before, during and after bath application of NMDA and 5-HT (Fig. 5) Respiratory frequency changes during drug and washout conditions were normalized and expressed as a percent of control values A grand mean ± SD across n animals was calculated Further information on research design is available in the Nature Portfolio Reporting Summary linked to this article The data that support the findings can be found in the source data provided with the paper. Original microscopy data have been deposited to Mendeley Data (https://doi.org/10.17632/wpbyxgfr96.1). For other raw data, example files can be obtained upon request. Source data are provided with this paper A review of the control of breathing during exercise The interactions between locomotion and respiration Absent phasing of respiratory and locomotor rhythms in running mice Respiratory and stepping frequencies in conscious exercising cats Cardiovascular and ventilatory responses to dynamic exercise during epidural anaesthesia in man Central activation of autonomic effectors during mental simulation of motor actions in man Breathing pattern and metabolic behavior during anticipation of exercise Identification of higher brain centres that may encode the cardiorespiratory response to exercise in humans Stimulation by central command of locomotion respiration and circulation during exercise Exercise hyperpnea and locomotion: parallel activation from the hypothalamus Chapter 4–supraspinal control of locomotion: the mesencephalic locomotor region Normal and pathological neuronal 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Modulation of respiratory activity by locomotion in lampreys Phox2b-expressing neurons of the parafacial region regulate breathing rate Photostimulation of retrotrapezoid nucleus phox2b-expressing neurons in vivo produces long-lasting activation of breathing in rats Respiratory-related activation of human abdominal muscles during exercise The effects of locomotion on respiratory muscle activity in the awake dog Differential contribution of the retrotrapezoid nucleus and C1 neurons to active expiration and arousal in rats Role of parafacial nuclei in control of breathing in adult rats Pontomedullary and hypothalamic distribution of Fos-like immunoreactive neurons after acute exercise in rats The role of parafacial neurons in the control of breathing during exercise Leptin action on GABAergic neurons prevents obesity and reduces inhibitory tone to POMC neurons A group of glutamatergic interneurons expressing high levels of both neurokinin-1 receptors and somatostatin identifies the 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modules for spatial orienting in the mouse superior colliculus Activation of groups of excitatory neurons in the mammalian spinal cord or hindbrain evokes locomotion Dual-mode operation of neuronal networks involved in left-right alternation Distribution of networks generating and coordinating locomotor activity in the neonatal rat spinal cord in vitro: a lesion study Periodic remodeling in a neural circuit governs timing of female sexual behavior AAV-mediated anterograde transsynaptic tagging: mapping corticocollicular input-defined neural pathways for defense behaviors Distinct rhythm generators for inspiration and expiration in the juvenile rat Breathing without CO(2) chemosensitivity in conditional Phox2b mutants The retrotrapezoid nucleus neurons expressing Atoh1 and Phox2b are essential for the respiratory response to CO(2) V2a neurons constrain extradiaphragmatic respiratory muscle activity at rest The peptidergic control circuit for sighing Brainstem projections to the major respiratory neuron populations in the medulla of the cat The role of the periaqueductal gray in escape behavior Descending command systems for the initiation of locomotion in mammals The multifunctional mesencephalic locomotor region Mesencephalic cuneiform nucleus and its ascending and descending projections serve stress-related cardiovascular responses in the rat Sciatic nerve stimulation activates the retrotrapezoid nucleus in anesthetized rats The rhythm section: an update on spinal interneurons setting the beat for mammalian locomotion A toolbox of Cre-dependent optogenetic transgenic mice for light-induced activation and silencing Expression pattern of a Krox-20/Cre knock-in allele in the developing hindbrain Principles for applying optogenetic tools derived from direct comparative analysis of microbial opsins Descending command neurons in the brainstem that halt locomotion Vertebral landmarks for the identification of spinal cord segments in the mouse Retrotrapezoid nucleus and parafacial respiratory group Afadin signaling at the spinal neuroepithelium regulates central canal formation and gait selection Download references Present address: Institute of Functional Genomics Institut de Biologie de l’École Normale Supérieure (IBENS) performed and analyzed all experiments with contributions from S.D contributed to histology experiments for the revision work prepared figures with contributions from J.B The authors declare no competing interests Nature Communications thanks Alexander Gourine reviewer(s) for their contribution to the peer review of this work Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations Download citation DOI: https://doi.org/10.1038/s41467-023-38583-6 Anyone you share the following link with will be able to read this content: Sorry, a shareable link is not currently available for this article. Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily. 28-year-old Yannis Cleymans from Herent (Flemish Brabant) has discovered a beached whale on the east coast of Canada thanks to his use of Google Maps. The Fleming writes articles about eastern Canada on Wikipedia as a hobby and stumbled upon the remarkable satellite image during his research. ‘I was expecting 10 likes, and suddenly I got a call from CBC,’ he says. ‘It’s a hobby, I spend an hour every day writing articles on Wikipedia,’ he explains. ‘I have always had a fascination with geography. As a child, I lived in the United States due to my father's work, but I felt especially connected to eastern Canada. After all, that is on the same latitude as Belgium.’ ‘Back home in Belgium, every day I write about Canada’s villages and rivers.’ While researching an article, Yannis came across a remarkable satellite image. ‘Before I start writing, I always look at the area via Google Maps. While studying the remote inlet of Clay Cove, I saw the whale. I didn't believe my eyes and posted the discovery in a Facebook group for beachcombers. I was expecting 10 likes at most, but suddenly the post went viral.’ With more than 5,000 likes, the post also caught the eye of a local journalist. Yannis made it onto the Canadian news and even made his debut on Canadian radio. ‘I didn't expect that at all,’ Yannis says. ‘But a Belgian discovering something like that, almost 4,000 kilometres away: they thought that was quite something.’ The satellite images turned out to have been made two years ago. ‘A few months after the sperm whale washed ashore, someone already bumped into the animal. At that time only some rotting blubber remained. Another year later, someone found some more bones and teeth. Being such a remote village, the news never went far. After my story was broadcast, another Canadian went to have a look, but found nothing left. Nature had already done its work.’ ‘Finding the whale was a fun experience. I am curious to see what reactions I will get from Canadians living in the area.’ Developing IQVIA’s positions on key trends in the pharma and life sciences industries, with a focus on EMEA. IQVIA is a world leader in using data, technology, advanced analytics and human expertise. We understand that part of your role involves spending a lot of time on the road, which is why at IQVIA we give you the opportunity to schedule your own agenda. Join IQVIA Belux and see where your skills can take you. "Join IQVIA Belux and see where your skills can take you. At IQVIA Belux, we are driving healthcare forward together. We are united as one team who share many different personalities, stories and experiences, but ultimately, who share a passion for healthcare. We are IQVIA. Partner and innovate to drive healthcare forward and make better decisions based on factual insights. 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Text description provided by the architects Warot Building is the latest addition to the public facilities of the village of Winksele It hosts a wide range of activities in a carefully designed multifunctional hall The building is situated on an existing sports campus that is surrounded by rural landscape and public facilities The large bay window and the covered outdoor space activate the adjoining fields for play The intervention is made up of a new building and a playing field, separated by a brook which will soon be crossed by a new bridge by UTIL. The bridge is positioned in such a way that it creates a new, soft connection between the existing sports facilities and the local school, child care, and youth centre across the brook. A large bay window with a single fold emphasises the new connection with the village and offers it a welcoming facade. AxonometricCommon materials were chosen for their simplicity, ease of construction, durability, and ease of maintenance while assuring a low ecological impact.  Sand-coloured brick, brown concrete flooring, steel, and metal ceilings are brought together through sensitive detailing to elevate their utilitarian nature. © Jeroen VerrechtThe clear and symmetrical plan ensures that the cost of the structure is reduced. The steel beams of the roof structure support a steel perforated corrugated sheet. The choice to perforate this structural sheet and keep it exposed ensures the acoustic quality inside the building while avoiding the cost of expensive acoustic finishing. You'll now receive updates based on what you follow Personalize your stream and start following your favorite authors If you have done all of this and still can't find the email The most-read stories on IEEE Spectrum right now You can read this article in 1 minuteAgnieszka Kulikowska - Wielgus Four Belgian municipalities have introduced traffic restrictions on local roads heavy goods transit can only use the main roads Photo credits @ Wikipedia Commons/Bjung/CC BY-SA 3.0 the Belgian municipalities of Steenokkerzeel located in the province of Flemish Brabant east of Brussels have implemented a common route system for HGV transit traffic This involves the introduction of a tonnage restriction of up to 3.5 tonnes as reported by the Belgian hauliers’ organisation Febetra transit traffic of heavy goods vehicles will only be permitted on the following main roads: The new system applies only to vehicles intended for the carriage of goods in transit deliveries) remains permitted at all times Pölös Zsófia Journalist Trans.info | 5.05.2025 Agnieszka Kulikowska - Wielgus Journalist Trans.info | 5.05.2025 Sabina Koll Journalist Trans.info | 5.05.2025 GXO to manage Northern Italy transport operations for PRG retail groupPölös Zsófia Journalist Trans.info | 5.05.2025 Trina and Jaydon Roberts recently competed in the Icenation Online Belgium Archery Shoot with archers from across the globe competing Both competed in the Recurve – 18m WA Division Jaydon Roberts finished second with a score of 540 points He finished nine points behind the winner Vincent Herent from France and was 12 points ahead of the third place finisher Alain Daniel from Switzerland Jaydon Roberts shot the highest with a score of 275; he then shot a second round 265 Trina Roberts finished sixth with a score of 439 which was the seventh best in the round; she then shot 219 #Archery Category: All, Sports Lode Dubois from Herent (Flemish Brabant) and his friends have been fined for excessive speed on ski slopes in the Italian resort of Livigno Italian police maintain the Flemish skier raced down the slopes at over 100 kilometres per hour It was in early January that Lode Dubois headed for the slopes of Livigno in Italy for a week’s skiing but when we came down the mountain on the third or fourth day of our trip we were stopped by the Italian police," Lode relates "They told us we were skiing way too fast as much as 100 kilometres per hour according to the police we were fined for misbehaviour on the ski slope." Lode and his friends had to hand in their ski passes They also had to accompany officers to the police station to make a statement I had surgery on my back less than a year ago and have to be careful not to fall at high speed but this did put a damper on my holiday spirit!" Please enable JS and disable any ad blocker